Abstract
Water proton T 1 –1 measurements at magnetic fields between 0.01 and 50 MHz [nuclear magnetic relaxation dispersion (NMRD) measurements] have been performed on solutions of phthalate dioxygenase (PDO) reconstituted at the catalytic iron site with copper(II) or manganese(II). The data show evidence of a weakly coordinated water molecule in CuPDO; in the presence of the substrate, phthalate, this water appears to become even less tightly bound, and an additional tightly coordinated water can be detected. In PDO reconstituted with manganese, one tightly coordinated water is detected in the presence and in the absence of phthalate. An attempt is made to reconcile these data with low-temperature near-IR magnetic circular dichroism and X-ray absorption data, which show that PDO reconstituted with iron or cobalt is six-coordinate in the absence of substrate and five-coordinate in the presence of substrate.
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Received: 24 April 1996 / Accepted: 17 July 1996
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Bertini, I., Luchinat, C., Mincione, G. et al. NMRD studies on phthalate dioxygenase: evidence for displacement of water on binding substrate. JBIC 1, 468–475 (1996). https://doi.org/10.1007/s007750050080
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DOI: https://doi.org/10.1007/s007750050080